Planting guide

Check seed stocks before planting - some germinate better than others!!

Hydroponics video: https://www.youtube.com/watch?v=c9neVLaS63c

Hydroponics paper: http://www.plantmethods.com/content/9/1/4

2012 Schedule

Potting on soil

date wks before Lab purpose per pot pots/pair # pots
1/4 8 0.3 demo 3-5 1 10
1/11 7 0.3 demo 3-5 1 10
3 1.1 DNA lots! 4 40
1/18 6 0.3 demo 3-5 1 10
2 1.1 DNA lots! 4 40
1/25 5 0.3 demo 3-5 1 10
2/1 4 0.3 demo 3-5 1 10
2/8 3 0.3 demo 3-5 1 10
2/15 2 0.3 demo 3-5 1 10
2/27 6 5.3 projects 3-5 1 10
3/6 5 5.3 projects 3-5 1 10
3/13 4 5.3 projects 3-5 1 10
3/20 3 5.3 projects 3-5 1 10
3/27 2 5.3 projects 3-5 1 10

Planting on plates

date for Lab purpose # plates notes
2/22 0.3 demo 10 1 weeks before lab
3/26 5.3 projects 10 2 weeks before lab
4/3 5.3 projects 10 1 weeks before lab
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Calendar Spring 2012

January 2012

Sun Mon Tue Wed Thurs Fri Sat
1 Holiday 2 3 4 Plant 5 6 7
8 9 10 11 Plant 12 13 14
15 16 Holiday 17 18 Plant 19 20 21
22 23 24 25 Plant 26 27 28
29 30 31


February 2012

Sun Mon Tue Wed Thurs Fri Sat
1 Plant 2 3 4
5 6 7 8 Plant 9 10 11
12 13 14 15 Plant 16 17 18
19 20 Holiday 21 22 23 24 25
26 27 Plant 28 29


March 2012

Sun Mon Tue Wed Thurs Fri Sat
1 2 3
4 5 6 Plant 7 8 9 10
11 12 13 Plant 14 15 16 17 Spring Break
18 19 20 Plant 21 22 23 24
25 26 27 Plant 28 29 30 31


April 2012

Sun Mon Tue Wed Thurs Fri Sat
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 Holiday 17 (Mon) 18 19 20 21
22 23 24 25 26 27 28
29 30


May 2012

Sun Mon Tue Wed Thurs Fri Sat
1 Last day 2 3 4 5
6 7 8 9 10 11 12
13 14 15 Grades due 16 17 18 19
20 21 22 23 24 25 26
27 28 Holiday 29 30 31

On plates

Planting on Plates

Make MS plates: http://wahoo.nsm.umass.edu/content/ms-plates

  • To get weakling mutants started before potting in soil
  • Or to plant seeds on specified media
  • Or to see roots

Seed Preparation

  • Put appropriate number of seeds in a sterile microfuge tube
  • Sterilize with EtOH (or sterilize with bleach – see below)
  • Sterilize filter paper with EtOH, let dry in sterile hood (can set it on top of an open sterile petri plate cover
  • Add 95% EtOH to seeds for 5 min, dump seeds onto filter paper
  • let dry, tap onto agar
  • Make a line of seeds in agarose above the midline of the plate:
  • Tape plates with Micropore surgical tape
  • Refrigerate the plates for 2 days
  • Move plates to growth chamber
  • Stand plates on edge under the lights. Tape several together.

OR Sterilize with bleach solution

  • Add ~1mL of bleach solution to each tube
  • Cap and shake
  • Leave tube on its side for 20-30 min, shaking periodically
  • Put upright after last shake (so seeds settle)
  • In laminar flow hood
    • Use a pipetman that has been sprayed with ethanol and air-dried in the hood
    • Remove as much bleach solution as possible from the tube
    • Add ~1 mL sterile ddH2O, mix and let seeds settle again
    • Do at least 4 washes, to dilute out the bleach
    • Add ~1mL sterile 0.1% agarose (so seeds will be suspended in the liquid)

Bleach solution for sterilizing seeds

  • 7 mL sterile ddH2O (must be freshly made!)
  • 3 mL Chlorox
  • 1 uL Triton X-100 (detergent to help get bleach into seed crevices)

0.1% agarose for plating seeds

  • 100 mL sterile ddH2O
  • 0.1 g agarose . autoclave to melt; shake bottle after autoclaving

On soil

Arabidopsis planting guide - Plants on soil

Routine weekly planting to produce plant material for observation or DNA extraction

  • Pots
    • 6 pots in a big flat with no holes
    • fill with clump-free potting soil, leaving some space at the rim
  • Treat with Gnatrol for fungus gnats
    • saturate with hot water the night before Teddi comes
    • Arrange for Teddi to do Gnatrol treatment
  • OR Heat in oven
    • pre-heat oven to 225F (=~107C)
    • Put soil-filled pots in aluminum lasagne pan
    • Cover with foil
    • insert thermometer in one pot near center
    • Heat to 180F (=~82C)
    • leave in oven for 30 min.
    • leave covered until ready to plant
  • Seeds for DNA
    • Start a set 4 weeks before needed, another set 3 weeks before
    • Columbia wild type (col-0)
    • Scatter seeds on surface of soil (remoisten before if it is dry)
    • Put on plastic cover
    • Put in refrigerator, keep dark (this synchronizes germination)
    • Transfer to growth chamber on the third day
    • If no room in fridge, put seeds in microfuge tube, add water, refrigerate in dark 2 days, then plant
  • Seeds for demo or experimental plants
    • Put seeds in microfuge tube, water, refrigerate, keep dark
    • 3rd day: pour into small beaker, add more water
    • Deposit one seed to each corner of the pot, plus one in the middle with a transfer pipet.
    • Transfer to growth chamber
  • Growth Chamber
    • 22C, 16H light, schedule continuous program
    • Water bottom flat ~3x per week
    • Remove cover after plants are well established.

hydroponics

http://www.plantmethods.com/content/9/1/4

http://www.bio-world.com/productinfo/3_44_292/124429/Magenta-GA-Plant-Cu...

2015 attempt

  • Make 1x MS, pH to 5.6

  • Split: most as growth medium, a little bit for 0.7% agar (err on the low end), sterilize

  • Cut lids off microfuge tubes, poke a hole in each, sterilize in a beaker

  • line lids up, flat side down, on a strip of scotch tape

  • fill with 0.7% agar.

  • put lids into floatie or other rack.

  • put 1 sterilized seed in each hole. (in 0.1% agar)

  • Wrap with saran wrap

  • Cover tightly with foil, stratify in refrigerator for 2-3 days

  • Remove foil, put container in growth chamber at 16h day, 22C

  • Transfer lid to 50 mL tube with hole drilled in it at ~3 weeks.

  • Aerate or stir medium